Difference between revisions of "Part:BBa K801073"
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===Cloning into pSB1C3=== | ===Cloning into pSB1C3=== | ||
| − | The cloning into pSB1C3 was proved by performing an analytical digest. | + | The cloning into pSB1C3 was proved by performing an analytical digest with XbaI and PstI. |
[[Image:TUM12_analyticaldigestCaXMT1cassette.jpg|100px]] | [[Image:TUM12_analyticaldigestCaXMT1cassette.jpg|100px]] | ||
Revision as of 12:51, 3 October 2012
CaXMT1 expression cassette for yeast
Yeast expression cassette for CaXMT1 (BBa_K801070) controlled by the yeast TEF2 promoter (BBa_K801010) and the yeast ADH1 terminator (BBa_K801012).
This BioBrick is the generator for the enzyme xanthosine N-methyltransferase 1 (CaXMT1) and part of the caffeine synthesis pathway. For creating an expression cassette with all three enzymes of the caffeine synthesis pathway BBa_K801077 based on the substrate Xanthosine different promoters and terminators were assembled to each enzyme. CaXMT1 is regulated by the constitutive promoter Tef2, which is a strong yeast promoter. The used terminator Adh1, is a widely used yeast terminator. The Tef2 promoter was prefered compared to the Tef1 promoter (which is even stronger) in order to limit metabolic stress, which could result in a positive selection of natural mutants (with regard to genome integration).
Cloning into pSB1C3
The cloning into pSB1C3 was proved by performing an analytical digest with XbaI and PstI.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 138
- 1000COMPATIBLE WITH RFC[1000]